The CD16
+
Monocyte Isolation Kit has been developed for a two-step isolation of the CD14
+
CD16
+
monocyte subset from PBMCs.

Data and images for
CD16
+
Monocyte Isolation Kit
, human

Figures

Figure 1

CD16
+
monocytes were isolated from PBMCs using the CD16
+
Monocyte Isolation Kit. In a first step, granulocytes and NK cells were depleted. In a second step, CD16
+
cells were enriched. An LD Column and a MidiMACS™ Separator were used for the depletion step, and an MS Column and a MiniMACS™ Separator for the positive selection step.
PBMCs before separation
Pre-enriched CD16
+
monocytes
View details

Figure 1

CD16
+
monocytes were isolated from PBMCs using the CD16
+
Monocyte Isolation Kit. In a first step, granulocytes and NK cells were depleted. In a second step, CD16
+
cells were enriched. An LD Column and a MidiMACS™ Separator were used for the depletion step, and an MS Column and a MiniMACS™ Separator for the positive selection step.
View details

Figure 1

CD16
+
monocytes were isolated from PBMCs using the CD16
+
Monocyte Isolation Kit. In a first step, granulocytes and NK cells were depleted. In a second step, CD16
+
cells were enriched. An LD Column and a MidiMACS™ Separator were used for the depletion step, and an MS Column and a MiniMACS™ Separator for the positive selection step.
Enriched CD16
+
monocytes
View details

Figure 1

CD16
+
monocytes were isolated from PBMCs using the CD16
+
Monocyte Isolation Kit. In a first step, granulocytes and NK cells were depleted. In a second step, CD16
+
cells were enriched. An LD Column and a MidiMACS™ Separator were used for the depletion step, and an MS Column and a MiniMACS™ Separator for the positive selection step.
View details

Figure 1

CD16
+
monocytes were isolated from PBMCs using the CD16
+
Monocyte Isolation Kit. In a first step, granulocytes and NK cells were depleted. In a second step, CD16
+
cells were enriched. An LD Column and a MidiMACS™ Separator were used for the depletion step, and an MS Column and a MiniMACS™ Separator for the positive selection step.

Figure 2

View details
Working scheme for the isolation of CD16
+
monocytes from PBMCs.

Figure 2

Working scheme for the isolation of CD16
+
monocytes from PBMCs.

Specifications for
CD16
+
Monocyte Isolation Kit
, human

Overview

The CD16
+
Monocyte Isolation Kit has been developed for a two-step isolation of the CD14
+
CD16
+
monocyte subset from PBMCs.

Detailed product information

Background information

Intermediate and non-classical monocytes together are referred to as CD16
+
monocytes
1
and account for about 5–10% of all blood monocytes. CD14
+
CD16
+
monocytes differ from CD14
+
CD16
-
monocytes with regard to phenotype and immunological function. CD16
+
monocytes are, for example, considered more mature and are believed to have a higher T cell stimulatory capacity than CD16
-
monocytes.
2, 3

Detailed separation procedure

Isolation of CD16
+
monocytes is performed in a two-step procedure. Prior to separation, cells are incubated with FcR Blocking Reagent. Granulocytes and NK cells are then magnetically labeled with a cocktail of CD15 MicroBeads and CD56 MicroBeads, and depleted over a MACS Column. The flow-through in this first separation step contains pre-enriched, unlabeled CD16
+
monocytes. In the second separation step, CD16
+
monocytes are efficiently isolated by positive selection after labeling with CD16 MicroBeads.

Downstream applications

Isolated blood CD16
+
monocytes are suitable, for example, for investigations on CD16
+
monocyte maturation, migration, and differentiation, or for studies on antigen uptake, antigen processing and presentation to T cells.

Columns

For the first magnetic separation (depletion): LD or autoMACS
®
Columns. For the second magnetic separation (positive selection): MS, LS, or autoMACS Columns.

References for
CD16
+
Monocyte Isolation Kit
, human

Publications

  1. Ziegler-Heitbrock et al. (2010) Nomenclature of monocytes and dendritic cells in blood. Blood 116: e74-80
  2. Ziegler-Heitbrock, H. W. (1996)
    Heterogeneity of human blood monocytes: the CD14
    +
    CD16
    +
    subpopulation.
    Immunol. Today 17: 424-428
  3. Grage-Griebnow, E. et al. (2001) Heterogeneity of human peripheral blood monocyte subsets. J. Leukoc. Biol. 69: 11-20

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